Human tissue samples were obtained from several sources: Clinomics Biosciences (Pittsfield, MA), Clontech, AllCells (Berkeley, CA), Clonetics/BioWhittaker (Walk-ersville, MD), AMS Biotechnology (Abingdon, Oxfordshire, U.K.), and the University of California at San Diego. When samples from four or more subjects were available, equal numbers of male and female subjects were used to make two independent pools; when fewer than four samples were available, RNA samples were pooled, and duplicate amplifications were performed for each pool (detailed annotation for human samples is on our web site, http:\\symatlas.gnf.org) Extracted RNA was additionally purified on RNeasy mini columns (Qiagen). RNA quantity and quality was verified on Bioanalyser 2100 (Agilent).
Five micrograms of total RNA was used to synthesize cDNA that was then used as a template to generate biotinylated cRNA using Enzo kits.
standard Affymetrix procedures except for use of a custom 24-channel fluidics machine and GCS2500 scanner